Endotoxin contamination of different plasmid preparation methods
Influence of endotoxins on biological applications
Removal of endotoxins
Endotoxin-free plasticware and glassware
To avoid recontamination of plasmid DNA after initial endotoxin removal, we recommend using only new plasticware that is certified to be pyrogen- or endotoxin-free. Endotoxin-free or pyrogen-free plasticware can be obtained from many different suppliers. Endotoxins adhere strongly to glassware and are difficult to remove completely during washing. Standard laboratory autoclaving procedures have little or no effect on endotoxin levels. Moreover, if the autoclave has previously been used for bacteria, the glassware will become extensively contaminated with endotoxin molecules. Heating glassware at 180°C overnight is recommended to destroy any attached endotoxin molecules. For further reading on endotoxin removal, please refer to the appropriate literature. It is also important not to recontaminate the purified endotoxin-free DNA by using reagents that are not endotoxin-free. All buffers supplied with the EndoFree Plasmid Kits are tested and certified to be endotoxin-free, as are the water for preparation of 70% ethanol and the TE buffer for resuspension.